| 转基因番茄防龋疫苗中外源目的基因拷贝数的检测 |
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| 引用本文： | 白国辉,刘建国,田源,陈筑,白朋元,韩琪,顾瑜,管晓燕,王海慧. 转基因番茄防龋疫苗中外源目的基因拷贝数的检测[J]. 上海口腔医学, 2013, 22(6): 613-617 |
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| 作者姓名： | 白国辉 刘建国 田源 陈筑 白朋元 韩琪 顾瑜 管晓燕 王海慧 |
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| 作者单位： | (1.遵义医学院 贵州省高等学校口腔疾病研究特色重点实验室，2.医学与生物学研究中心，贵州 遵义 563000；3.贵阳市口腔医院，贵州 贵阳 550002) |
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| 基金项目： | 国家自然科学基金(30160086，81260164)；贵州省科学技术基金[黔科合J字LKZ[2011]41号]；贵州省科技创新团队资助项目、贵州省重点学科建设资助项目；遵义医学院优秀科研团队培育项目[[2012]12号] |
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| 摘 要： | 目的：采用SYBR Green实时荧光定量 PCR方法检测转基因番茄防龋疫苗中外源目的基因的拷贝数。方法：用本课题组前期构建的重组质粒pEAC10、pEPC10作为标准品，SYBR Green实时荧光定量PCR法对含外源目的基因pacA-ctxB、pacP-ctxB的转基因番茄植株总基因组样本重复检测，取平均值作为目的基因拷贝数。结果：含pacA-ctxB转基因番茄植株外源目的基因的拷贝数为1.3，含pacP-ctxB转基因番茄植株外源目的基因的拷贝数为3.2。结论：构建的转基因番茄植株属低拷贝转基因植物，具有较好的稳定性。
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| 关 键 词： | 转基因番茄 实时荧光定量PCR SYBR Green 拷贝数 Ct 值 |
| 收稿时间： | 2013-04-05 |
| 修稿时间： | 2013-05-10 |
Detection of the exogenous gene copy number of the transgenic tomato anti-caries vaccine |
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| BAI Guo-hui,LIU Jian-guo,TIAN Yuan,CHEN Zhu,BAI Peng-yuan,HAN Qi,GU Yu,GUAN Xiao-yan,WANG Hai-hui. Detection of the exogenous gene copy number of the transgenic tomato anti-caries vaccine[J]. Shanghai journal of stomatology, 2013, 22(6): 613-617 |
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| Authors: | BAI Guo-hui LIU Jian-guo TIAN Yuan CHEN Zhu BAI Peng-yuan HAN Qi GU Yu GUAN Xiao-yan WANG Hai-hui |
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| Affiliation: | 1. The Special Key Laboratory of Oral Diseases Research, Higher Education Institution of Guizhou Province, 2. Research Center for Medicine & Biology, Zunyi Medical College. Zunyi 563000; 3. The Stomatological Hospital of Guiyang City. Guiyang 550002, Guizhou Province, China |
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| Abstract: | PURPOSE: To detect the exogenous gene copy number of the transgenic tomato anti-caries vaccine by using the SYBR Green real-time PCR. METHODS: Recombinant plasmid pEAC10 and pEPC10 were used as standard to detect genome samples of exogenous gene pacA-ctxB and pacP-ctxB by SYBR green fluorescent quantitation, then the average value was calculated as gene copy number. RESULTS: The copy number of the transgenic tomato carrying pacA-ctxB was 1.3 and the pacP-ctxB was 3.2. CONCLUSIONS: The transgenic tomato plants which have high stability are low-copy transgenic plants. |
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| Keywords: | Transgenic tomato Real-time fluorescence quantitative PCR SYBR Green Threshold cycle Copy number |
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